Details
| Translated title of the contribution | Identifizierung von RNA-Basenpaaren und vollständige Zuordnung von Nukleobasen-Resonanzen durch Protonen-detektierte Festkörper-NMR-Spektroskopie bei MAS Geschwindigkeiten von 100 kHz |
|---|---|
| Original language | English |
| Pages (from-to) | 23903-23910 |
| Number of pages | 8 |
| Journal | Angewandte Chemie - International Edition |
| Volume | 60 |
| Issue number | 44 |
| Early online date | 28 Sept 2021 |
| Publication status | Published - 25 Oct 2021 |
Abstract
Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional 13C- and 15N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using 1H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.
Keywords
- H detection, base-pair pattern, RNA structure, RNA-protein complex, solid-state NMR spectroscopy
ASJC Scopus subject areas
- Chemical Engineering(all)
- Catalysis
- Chemistry(all)
- General Chemistry
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In: Angewandte Chemie - International Edition, Vol. 60, No. 44, 25.10.2021, p. 23903-23910.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Identification of RNA Base Pairs and Complete Assignment of Nucleobase Resonances by Proton-Detected Solid-State NMR Spectroscopy at 100 kHz MAS
AU - Aguion, Philipp Innig
AU - Kirkpatrick, John
AU - Carlomagno, Teresa
AU - Marchanka, Alexander
N1 - Funding Information: This work was supported by the Deutsche Forschungsgemeinschaft (DFG grant CA294/21‐1 to TC). Open Access funding enabled and organized by Projekt DEAL.
PY - 2021/10/25
Y1 - 2021/10/25
N2 - Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional 13C- and 15N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using 1H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.
AB - Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional 13C- and 15N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using 1H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.
KW - H detection
KW - base-pair pattern
KW - RNA structure
KW - RNA-protein complex
KW - solid-state NMR spectroscopy
UR - http://www.scopus.com/inward/record.url?scp=85115880766&partnerID=8YFLogxK
U2 - 10.1002/anie.202107263
DO - 10.1002/anie.202107263
M3 - Article
C2 - 34379871
AN - SCOPUS:85115880766
VL - 60
SP - 23903
EP - 23910
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
SN - 1433-7851
IS - 44
ER -